TY - JOUR
T1 - Detection of LacZ-Positive Cells in Living Tissue with Single-Cell Resolution
AU - Doura, Tomohiro
AU - Kamiya, Mako
AU - Obata, Fumiaki
AU - Yamaguchi, Yoshifumi
AU - Hiyama, Takeshi Y.
AU - Matsuda, Takashi
AU - Fukamizu, Akiyoshi
AU - Noda, Masaharu
AU - Miura, Masayuki
AU - Urano, Yasuteru
N1 - Funding Information:
This research was supported in part by the Ministry of Education, Culture, Sports, Science and Technology of Japan (Grant-in-Aid for Scientific Research (KAKENHI), grants 26111012 and 16H02606 to Y.U., grant 15H05951 to M.K., grants 26111726 and 26293043 to T.Y.H., grants by the Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS) to Y.U.), by JSPS Core-to-Core Program, A. Advanced Research Networks (to Y.U.), by The Daiichi-Sankyo Foundation of Life Science (grant to Y.U.), by Japan foundation for applied enzymology (to M.K.). We thank Dr. Kazunari Hisadome for helpful discussions.
Publisher Copyright:
© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
PY - 2016/8/8
Y1 - 2016/8/8
N2 - The LacZ gene, which encodes Escherichia coli β-galactosidase, is widely used as a marker for cells with targeted gene expression or disruption. However, it has been difficult to detect lacZ-positive cells in living organisms or tissues at single-cell resolution, limiting the utility of existing lacZ reporters. Herein we present a newly developed fluorogenic β-galactosidase substrate suitable for labeling live cells in culture, as well as in living tissues. This precisely functionalized fluorescent probe exhibited dramatic activation of fluorescence upon reaction with the enzyme, remained inside cells by anchoring itself to intracellular proteins, and provided single-cell resolution. Neurons labeled with this probe preserved spontaneous firing, which was enhanced by application of ligands of receptors expressed in the cells, suggesting that this probe would be applicable to investigate functions of targeted cells in living tissues and organisms.
AB - The LacZ gene, which encodes Escherichia coli β-galactosidase, is widely used as a marker for cells with targeted gene expression or disruption. However, it has been difficult to detect lacZ-positive cells in living organisms or tissues at single-cell resolution, limiting the utility of existing lacZ reporters. Herein we present a newly developed fluorogenic β-galactosidase substrate suitable for labeling live cells in culture, as well as in living tissues. This precisely functionalized fluorescent probe exhibited dramatic activation of fluorescence upon reaction with the enzyme, remained inside cells by anchoring itself to intracellular proteins, and provided single-cell resolution. Neurons labeled with this probe preserved spontaneous firing, which was enhanced by application of ligands of receptors expressed in the cells, suggesting that this probe would be applicable to investigate functions of targeted cells in living tissues and organisms.
KW - fluorescent probes
KW - lacZ
KW - quinone methide intermediates
KW - single-cell resolution
KW - β-galactosidase
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U2 - 10.1002/anie.201603328
DO - 10.1002/anie.201603328
M3 - Article
C2 - 27400827
AN - SCOPUS:84978245551
SN - 1433-7851
VL - 55
SP - 9620
EP - 9624
JO - Angewandte Chemie - International Edition
JF - Angewandte Chemie - International Edition
IS - 33
ER -